Gly-Arg-AMC (hydrochloride) (Synonyms: Gly-Arg-7-amino-4-methylcoumarin, GR-AMC, GR-7-amino-4-methylcoumarin) |
| Catalog No.GC45668 |
Gly-Arg-AMC (hydrochloride) is a fluorescent substrate for Cathepsin C.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 70274-78-1
Sample solution is provided at 25 µL, 10mM.
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Gly-Arg-AMC (hydrochloride) is a fluorescent substrate for Cathepsin C. Upon hydrolysis by Cathepsin C, Gly-Arg-AMC (hydrochloride) is cleaved into Gly (glycine) and Arg-AMC (arginine-7-amino-4-methylcoumarin), with AMC emitting blue-green fluorescence. Gly-Arg-AMC (hydrochloride) can be used to detect the activity of Cathepsin C (excitation/emission: 360/460nm)[1].
References:
[1] Rubach JK, Cui G, Schneck JL, Taylor AN, Zhao B, Smallwood A, Nevins N, Wisnoski D, Thrall SH, Meek TD. The amino-acid substituents of dipeptide substrates of cathepsin C can determine the rate-limiting steps of catalysis. Biochemistry. 2012 Sep 25;51(38):7551-68.
1. Buffer Preparation
Prepare a 10μL reaction mixture composed of 50mM sodium acetate (pH 5.5), 30mM NaCl, 1mM EDTA, 1mM CHAPS, 1mM DTT (Buffer A), and 10% (v/v) DMSO.
2. Fluorescence Detection Conditions and Equipment
Peptidolysis is measured at 21°C in 384-well Greiner low-volume polypropylene plates. Fluorescence of the aminomethyl coumarin (AMC) product is measured using an Acquest fluorimeter with an excitation wavelength of 360nm, an emission wavelength of 460nm, and a 400nm cutoff filter.
3. Experimental Procedure
(1)Add 0.8μL of Gly-Arg-AMC (hydrochloride) to the 10μL reaction mixture.
(2)Initiate the reaction by adding Cathepsin C to achieve a concentration range of 0.01 - 100nM.
(3)Calculate the activity of Cathepsin C based on the fluorescence intensity.
This protocol only provides a guideline, and should be modified according to your specific needs
References:
[1] Rubach JK, Cui G, Schneck JL, Taylor AN, Zhao B, Smallwood A, Nevins N, Wisnoski D, Thrall SH, Meek TD. The amino-acid substituents of dipeptide substrates of cathepsin C can determine the rate-limiting steps of catalysis. Biochemistry. 2012 Sep 25;51(38):7551-68.
| Cas No. | 70274-78-1 | SDF | |
| Synonyms | Gly-Arg-7-amino-4-methylcoumarin, GR-AMC, GR-7-amino-4-methylcoumarin | ||
| Canonical SMILES | O=C1C=C(C)C2=C(C=C(NC([C@@H](NC(CN)=O)CCCNC(N)=N)=O)C=C2)O1 | ||
| Formula | C18H24N6O4.XHCl | M.Wt | 388.4 |
| Solubility | DMF: 30mg/mL,DMSO: 30mg/mL,DMSO:PBS (pH 7.2) (1:1): 0.5mg/mL,Ethanol: 20mg/mL | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 2.5747 mL | 12.8733 mL | 25.7467 mL |
| 5 mM | 514.9 μL | 2.5747 mL | 5.1493 mL |
| 10 mM | 257.5 μL | 1.2873 mL | 2.5747 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 2 reference(s) in Google Scholar.)
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